Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-01-08 to 2013-04-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name: Carbamic acid, N-(3-(triethoxysilyl)propyl)2-(ethyl-(4-nitrosophenyl) amino) ethyl ester
- Internal Code: SAT 120009
- Batch No. 12/34#
- CAS No.: 119231-94-7
- Purity: 92.8 wt% (NMR), 93.3 area% (HPLC)

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
HOUSING AND FEEDING CONDITIONS
- Full barrier in an air-conditioned room
- Temperature: 22 +/- 3 °C
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 0636)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups / individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 011012)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
cotton seed oil
Details on oral exposure:
The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals.
Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.
The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.
Doses:
The starting dose was selected to be 2000 mg/kg body weight. No compound-related mortality was recorded for any animal of step 1 or 2.
Based on these results and according to the acute toxic class method regime no further testing was required.
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
OBSERVATION PERIOD:
All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.

WEIGHT ASSESSMENT:
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

CLINICAL EXAMINATION:
A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention
given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for
clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central
nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor,
convulsions, salivation, diarrhoea, lethargy, sleep and coma.

PATHOLOGY:
At the end of the observation period all animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally
(Narcoren®, Merial; lot no.: 224052; expiry date: 05/2015) at a dosage of approx. 8 mL/kg bw.
All animals were subjected to gross necropsy. All gross pathological changes were recorded and in case of findings the tissues were preserved
for possible histopathological evaluation.
Statistics:
According to OECD guidelines, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation
of the results is not regarded as necessary.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
approximate LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
All animals survived until the end of the study showing signs of toxicity.
Clinical signs:
The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity,
moving of the bedding, recumbency, bradykinesia, kyphosis, piloerection, half eyelid closure and closed eyes. The clinical signs persisted
up to 3 days post-application.
Body weight:
Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain.
Gross pathology:
At necropsy, no macroscopic findings were observed in any animal of any step.

Any other information on results incl. tables

Table: Clinical Signs - Individual Data

Animal No.

/ Sex

Time of Observation Post-Dose

Observations

 

Step 1 (2000 mg/kg Body Weight)

1 / female

30 min

slightly reduced spontaneous activity, moving the bedding, partial recumbency, bradykinesia, half eyelid closure

1 h

slightly reduced spontaneous activity, moving the bedding, bradykinesia, kyphosis, slight piloerection, half eyelid closure

2 h

slightly reduced spontaneous activity, bradykinesia, kyphosis, moderate piloerection, half eyelid closure

3 h

moderately reduced spontaneous activity, bradykinesia, kyphosis, moderate piloerection, eyes closed

4 h

slightly reduced spontaneous activity, bradykinesia, slight piloerection

d 2

piloerection

d 3

piloerection

day 4 until the end of the observation period

no signs of toxicity

Animal No.

/ Sex

Time of Observation Post-Dose

Observations

 

Step 1 (2000 mg/kg Body Weight)

2 / female

30 min

slightly reduced spontaneous activity, moving the bedding, bradykinesia, slight piloerection

1 h

slightly reduced spontaneous activity, bradykinesia, slight piloerection

2 h

slightly reduced spontaneous activity, bradykinesia, slight piloerection, half eyelid closure

3 h

slight piloerection

4 h until the end of the observation procedure

no signs of toxicity

3 / female

30 min

slightly reduced spontaneous activity, moving the bedding, slight piloerection

1 h

slightly reduced spontaneous activity, bradykinesia, slight piloerection

2 h

slightly reduced spontaneous activity, bradykinesia, slight piloerection, half eyelid closure

3 h

slight piloerection

4 h until the end of the observation period

no signs of toxicity

Animal No.

/ Sex

Time of Observation Post-Dose

Observations

 

Step 2 (2000 mg/kg Body Weight)

4 / male

5 / male

6 / male

10 min

slightly reduced spontaneous activity, moving the bedding

30 min

moderately reduced spontaneous activity, moving the bedding, partial recumbency, bradykinesia, half eyelid closure

1 h

moderately reduced spontaneous activity, partial recumbency, bradykinesia, half eyelid closure

2 h, 3 h

moderately reduced spontaneous activity, half eyelid closure

4 h

slightly reduced spontaneous activity

day 2 until the end of the observation period

no signs of toxicity

d = day (day 1 = day of administration);           h = hour(s);          min = minute(s)

Table: Body Weight Development - Absolute Body Weights in g and Body Weight Gain in %

Animal No. / Sex

g

Day 1

g

Day 8

g

Day 15

%

Day 1-15

Step 1 (2000 mg/kg Body Weight)

1 / female

155

188

202

30

2 / female

177

204

220

24

3 / female

151

171

184

22

 

Animal No. / Sex

g

Day 1

g

Day 8

g

Day 15

%

Day 1-15

Step 2 (2000 mg/kg Body Weight)

4 / male

233

271

301

29

5 / male

239

285

318

33

6 / male

244

289

328

34

Table: Findings of Necropsy - Individual Data

Animal No.

/Sex

Organ

Macroscopic Findings

Step 1 (2000 mg/kg Body Weight)

1 / female

-

nsf

2 / female

-

nsf

3 / female

-

nsf

Step 2 (2000 mg/kg Body Weight)

4 / male

-

nsf

5 / male

-

nsf

6 /male

-

nsf

Table: LD50 Cut-Off

Dose (unit)

Number of Animals Investigated

Number of Intercurrent Deaths

LD50 Cut-Off

2000 mg/kg bw

6

0

5000 mg/kg bw

bw = body weight

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the present study, a single oral application of the test item SAT 120009 to rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity but no mortality.
The median lethal dose of SAT 120009 after a single oral administration to female and male rats, observed over a period of 14 days is:
LD50 (rat): > 2000 mg/kg bw
According to OECD Guideline 423, Annex 2 the LD50 cut-off (rat) under consideration of the clinical findings is 5000 mg/kg bw.
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test item SAT 120009 has no obligatory labelling requirement for toxicity.
According to Annex I of Regulation (EC) 1272/2008 (CLP-Regulation) the test item SAT 120009 has no obligatory labelling requirement for toxicity and is unclassified.
According to UN-GHS (Globally Harmonized Classification System) the test item SAT 120009 has obligatory labelling requirement for toxicity and is classified into Category 5.
Executive summary:

SUMMARY RESULTS

Two groups, one of three female and one of three male WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was suspended in the vehicle cottonseed oil at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

All animals used in the study after their entrance at BSL were allowed to acclimatise to the laboratory conditions for at least 5 days. The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

RESULTS PER STEP

Step

Sex/No.

Dose (mg/kg)

Number of Animals

Number of Intercurrent Deaths

1

female/1-3

2000

3

0

2

male/4-6

2000

3

0

All animals survived until the end of the study showing signs of toxicity.

The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, moving of the bedding, recumbency, bradykinesia, kyphosis, piloerection, half eyelid closure and closed eyes. The clinical signs persisted up to 3 days post-application.

Throughout the 14-day observation period, the body weight gain of the test animals was within the normal range of variation for this strain.

At necropsy, no macroscopic findings were observed in any animal of any step.


On the basis of the test results given below and in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC, the substance should be:

classified as very toxic

 

classified as toxic

 

classified as harmful

 

not classified

X

limit test

X

On the basis of the test results given below and in conformity with the criteria given inAnnex I of Regulation (EC) 1272/2008 (CLP-Regulation), the substance should be:

classified into category 1

 

classified into category 2

 

classified into category 3

 

classified into category 4

 

not classified

X

On the basis of the clinical findings and the test results given below and in conformity with the criteria given in UN-GHS (Globally Harmonized System of Classification and Labelling of Chemicals), the substance should be:

classified into category 1

 

classified into category 2

 

classified into category 3

 

classified into category 4

 

classified into category 5

X

not classified

 

LD50cut-off: 5000 mg/kg bw

LD50: > 2000 mg/kg bw

Species/strain: WISTAR Crl: WI(Han) rats

Number of animals: 3 per step / 2 steps performed

Vehicle: cottonseed oil

Method: OECD 423, Commission Regulation (EC) 440/2008, OPPTS 870.1000, OPPTS 870.1100

CONCLUSION

Under the conditions of the present study, a single oral application of the test item SAT 120009 to rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity but no mortality.

The median lethal dose of SAT 120009 after a single oral administration to female and male rats, observed over a period of 14 days is:

LD50 (rat): > 2000 mg/kg bw

According to OECD Guideline 423, Annex 2 the LD50 cut-off (rat) under consideration of the clinical findings is 5000 mg/kg bw.

In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC the test item SAT 120009 has no obligatory labelling requirement for toxicity.

According to Annex I of Regulation (EC) 1272/2008 (CLP-Regulation) the test item SAT 120009 has no obligatory labelling requirement for toxicity and is unclassified.

According to UN-GHS (Globally Harmonized Classification System) the test item SAT 120009 has obligatory labelling requirement for toxicity and is classified into Category 5.